Measurement of HIV Neutralization Using Quantitative Assays

Mascola, John R.

Abstract

Individual gp160 clones were screened for infectivity as Env-pseudotyped viruses in a luciferase reporter gene assay in JC53-BL (TZM-bl) cells. Functional env clones were sequenced and their neutralization phenotypes characterized by using soluble CD4, monoclonal antibodies and serum samples from infected individuals and non-infected recipients of a recombinant gp120 vaccine. Env clones from 12 R5 primary HIV-1 isolates were selected that were not unusually sensitive or resistant to neutralization and comprised a wide spectrum of genetic, antigenic and geographic diversity. These reference reagents will facilitate proficiency testing and other validation efforts aimed at improving assay performance across laboratories and can be used for standardized assessments of vaccine elicited neutralizing antibodies. An similar reference panel for clade C viruses has also been made.? ? Using the reference virus regents described above, the BSL3 core virology lab has screened hundreds of animal sera generated by investigators at the VRC.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Allergy and Infectious Diseases (NIAID)
• Type: Intramural Research (Z01)
• Project #: 1Z01AI005010-07
• Application #: 7732733
• Support Year: 7
• Fiscal Year: 2008
• Total Cost: $320,240

Institution

• Name: National Institute of Allergy and Infectious Diseases
• Country: United States

Publications

Li, Yuxing; Svehla, Krisha; Louder, Mark K et al. (2009) Analysis of neutralization specificities in polyclonal sera derived from human immunodeficiency virus type 1-infected individuals. J Virol 83:1045-59

Dey, Barna; Pancera, Marie; Svehla, Krisha et al. (2007) Characterization of human immunodeficiency virus type 1 monomeric and trimeric gp120 glycoproteins stabilized in the CD4-bound state: antigenicity, biophysics, and immunogenicity. J Virol 81:5579-93

Shu, Yuuei; Winfrey, Sarah; Yang, Zhi-Yong et al. (2007) Efficient protein boosting after plasmid DNA or recombinant adenovirus immunization with HIV-1 vaccine constructs. Vaccine 25:1398-408

Montefiori, David; Sattentau, Quentin; Flores, Jorge et al. (2007) Antibody-based HIV-1 vaccines: recent developments and future directions. PLoS Med 4:e348

Wang, Shixia; Pal, Ranajit; Mascola, John R et al. (2006) Polyvalent HIV-1 Env vaccine formulations delivered by the DNA priming plus protein boosting approach are effective in generating neutralizing antibodies against primary human immunodeficiency virus type 1 isolates from subtypes A, B, C, D and E. Virology 350:34-47

Rao, Srinivas S; Gomez, Phillip; Mascola, John R et al. (2006) Comparative evaluation of three different intramuscular delivery methods for DNA immunization in a nonhuman primate animal model. Vaccine 24:367-73

Wu, Lan; Yang, Zhi-Yong; Xu, Ling et al. (2006) Cross-clade recognition and neutralization by the V3 region from clade C human immunodeficiency virus-1 envelope. Vaccine 24:4995-5002

Pal, Ranajit; Kalyanaraman, V S; Nair, B C et al. (2006) Immunization of rhesus macaques with a polyvalent DNA prime/protein boost human immunodeficiency virus type 1 vaccine elicits protective antibody response against simian human immunodeficiency virus of R5 phenotype. Virology 348:341-53

Li, Y; Svehla, K; Mathy, N L et al. (2006) Characterization of antibody responses elicited by human immunodeficiency virus type 1 primary isolate trimeric and monomeric envelope glycoproteins in selected adjuvants. J Virol 80:1414-26

Louder, Mark K; Sambor, Anna; Chertova, Elena et al. (2005) HIV-1 envelope pseudotyped viral vectors and infectious molecular clones expressing the same envelope glycoprotein have a similar neutralization phenotype, but culture in peripheral blood mononuclear cells is associated with decreased neutralization sensi Virology 339:226-38